Keratinocyte Growth Kit (ATCC® PCS-200-040™)

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Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Product Format

frozen 1 Kit

Storage Conditions

-20°C (or -70°C for long-term storage)

Comments

The Keratinocyte Growth Kit contains components that when added to Dermal Cell Basal Medium (ATCC PCS-200-030) create a complete serum-free culture environment for keratinocytes. The complete keratinocyte growth medium is formulated to inhibit fibroblast growth, and the low calcium concentration (0.06 mM) slows differntiation. No feeder layers, extracellular matrix proteins or other substrares are required. The final concentration for each component in complete keratinocyte growth medium is as follows:

Extract P: 0.4%

rh TGF-alpha: 0.5 ng/mL

L-Glutamine: 6 mM

Hydrocortisone: 100 ng/mL

Insulin: 5 µg/mL

Epinephrine: 1.0 µM

Apo-transferrin: 5 µg/mL

Complete Growth Medium

Obtain one growth kit from the freezer; make sure that the caps of all components are tight.

Thaw the components of the growth kit just prior to adding to the basal medium. It is necessary to warm the L-glutamine component in a 37°C water bath and shake to dissolve any precipitates, prior to adding to the basal medium.

Decontaminate the external surfaces of all growth kit component vials and the basal medium bottle by spraying with 70% ethanol.

Using aseptic technique and working in a laminar flow hood or biosafety cabinet, transfer the indicated volume of each growth kit component to the bottle of basal medium using a separate sterile pipette for each.

Table 1. Keratinocyte Growth Kit Components

Component

Volume

Final Concentration

Extract P

2.0 mL

0.4%

rh TGF-a

0.5 mL

0.5 ng/mL

L-Glutamine

15.0 mL

6 mM

Hydrocortisone Hemisuccinate

0.5 mL

100 ng/mL

rh Insulin

0.5 mL

5 µg/mL

Epinephrine

0.5 mL

1.0 µM

Apo-Transferrin

0.5 mL

5 µg/mL

Antimicrobials and phenol red are not required for proliferation, but may be added if desired. The recommended volume of each optional component to be added to the complete growth media is summarized in Table 2.